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Freni was born into a working-class family in Modena; her mother and tenor Luciano Pavarotti's mother worked together and an aunt was the soprano Valentina Bartolomasi. She was a musically gifted child, and when 10 years old sang "Un bel dì vedremo" in a radio competition; the tenor Beniamino Gigli warned her, however, that she risked ruining her voice and advised her to give up singing until she was older. She resumed singing at the age of 17.

In 1978 she married Nicolai Ghiaurov, one of the leading operatic basses of the post-war period. Together they helped to establish the Centro Universale del Bel Canto in Vignola, where they began giving master classes in 2002. Following Ghiaurov's death in 2004, Freni continued their work of preserving the Bel Canto tradition, and currently teaches young singers from around the world.

Freni extended her repertoire and style during the 1990s with Italian Verismo, taking on the title roles of Cilea's Adriana Lecouvreur in Milan, Paris, Barcelona and New York, and Giordano's Fedora in London, Milan, New York, Torino, Barcelona and Zürich. In 1998, she performed Giordano's Madame Sans-Gêne in Catania. During this time she sang in the Russian operas of Tchaikovsky, appearing as Tatiana in Eugene Onegin, Lisa in The Queen of Spades, and Ioanna in The Maid of Orleans.